Entomopathogenic fungi detection and avoidance by mole crickets (Orthoptera: Gryllotalpidae)

A chamber to monitor mole cricket behavior was designed using two different soil-filled containers and photosensors constructed from infrared emitters and detectors. Mole crickets (Scapteriscus spp.) were introduced into a center tube that allowed them to choose whether to enter and tunnel in untreated soil or soil treated with Beauveria bassiana (Balsamo) Vuillemin. Each time the cricket passed through the photosensor located near the entrance of soil-filled containers, the infrared light was blocked and the exact moment that this occurred was logged onto a computer using custom-written software. Data examined included the first photosensor trigger, total number of sensor triggers, presence of tunneling, and final location of the cricket after 18 h. These behaviors were analyzed to discern differences in mole cricket behavior in the presence of different treatments and to elucidate the mechanism that mole crickets use to detect fungal pathogens. The first study examined substrate selection and tunneling behavior of the southern mole cricket, Scapteriscus borellii Giglio-Tos, to the presence of five strains of B. bassiana relative to a control. There were no differences between the first sensor trigger and total number of triggers, indicating the mole crickets are not capable of detecting B. bassiana at a distance of 8 cm. Changes in mole cricket tunneling and residence time in treated soil occurred for some strains of B. bassiana but not others. One of the strains associated with behavioral changes in the southern mole cricket was used in a second experiment testing behavioral responses of the tawny mole cricket, S. vicinus Scudder. In addition to the formulated product of this strain, the two separate components of that product (conidia and carrier) and bifenthrin, an insecticide commonly used to control mole crickets, were tested. There were no differences in mole cricket behavior between treatments in this study. The differences in behavioral responses between the two species could suggest a more sensitive chemosensory recognition system for southern mole crickets.     

Natural microbial control of crickets populations (Orthoptera: Gryllotalpidae: Scapteriscus borellii): regulation of populations aggregated in time and space

From 1983 through 1988, a total of 1,762 collections, containing 31,312 individuals of the mole cricket, Scapteriscus borellii, were made, principally in the State of S?o Paulo, Brazil. Collections were found to fit a negative binomial distribution both as whole and when divided into monthly collections. In these collections, an iridovirus, a entomogenous nematode, and the fungi Metarhizium anisopliae, Beauveria bassiana, Paecilomyces sp., and Entomophtora sp., were found to be agents of natural mortality, although usually as endozootics and relatively rarely as epizootics and panzootics. As a group, these diseases were also distributed in a binomial negative. These data suggest that the temporal and spatial aggregations of the mole crickets, produced by high rates of migration among suitable habitats, are adaptations to outbreaks of epidemics, which also serve as mole cricket population regulators. These ideas are develop and derived from simple mathematical models of population change.     

Effect of irrigation on the efficacy of insecticides for controlling two species of mole crickets (Orthoptera: Gryllotalpidae) on golf courses

Effects of irrigation regimen, quantity, and timing on the efficacy of three insecticides for controlling nymphs of the southern mole cricket, Scapteriscus borellii Giglio-Tos, and the tawny mole cricket, Scapteriscus vicinus Scudder, were studied on golf courses in 1997, 1998, and 1999. Two irrigation regimen tests using two rates of bifenthrin and lambda-cyhalothrin produced inconclusive results. Mole cricket damage ratings after the applications of bifenthrin (60 g [AI]/ha) and lambda-cyhalothrin (76 g [AI]/ha) were not significantly different among the four irrigation regimens (non-irrigation, irrigation before treatment, irrigation after treatment, and irrigation before and after treatment). Mole cricket damage rating after the application of bifenthrin (120 g [AI]/ha) under irrigation before and after irrigation was significantly better than those under other irrigation regimens at 14 and 21 d after treatment (DAT). Different irrigation quantity and irrigation timing (after insecticide treatment) did not significantly affect the performance of imidacloprid (434 g [AI]/ha) in the 1998 tests. However, the results from the 1999 test indicated that mole cricket damage ratings from the imidacloprid-treated plots were significantly different between 2 and 0.5 cm irrigation water after treatment at 21 and 28 DAT. Application of bifenthrin at a rate of 120 g (AI)/ha with 0.5 cm of irrigation water after treatment resulted in significantly lower mole cricket damage ratings than those of 1.0 and 2.0 cm of irrigation water after treatment at 30 DAT only in the 1998 test. Bifenthrin with irrigation at 1 h after insecticide treatment provided better mole cricket control than that of irrigation at 5 min after treatment at 30 DAT only in the 1998 test. Mole cricket damage ratings after application of bifenthrin were not significantly different between either irrigation quantity treatment or irrigation timing treatment in the 1999 tests. Possible effects of application timing, environmental conditions, irrigation practice, and insecticide physical properties on the results are discussed.     

Effect of combining imidacloprid and diatomaceous earth with Beauveria bassiana on mole cricket (Orthoptera: Gryllotalpidae) mortality

Sublethal doses of three orthopteran-derived strains of Beauveria bassiana (Balsamo) Vuillemin were topically applied to adult southern mole crickets, Scapteriscus borellii Giglio-Tos (Orthoptera: Gryllotalpidae), and tested in combination with substrate treatments of diatomaceous earth (DE) and imidacloprid. Crickets treated only with the high doses (10(8) conidia per cricket) of each of the three B. bassiana strains exhibited the shortest survival times as well as the highest percentage mortality at 28 d after treatment. However, these treatments did.not differ significantly from any of the diatomaceous earth combination treatments. Two of the strains tested, 5977 and 3622, exhibited synergistic interactions with DE, whereas the third strain, GHA, was not significant for synergy. Mortality caused by the combination treatment was still greater than the expected additive effect. DE abrades the insect cuticle and absorbs cuticular lipids, aiding the entry of germinating conidia into the mole cricket hemocoel. None of the three strains exhibited synergy when combined with imidacloprid, and mortality of all combination treatments was less than additive. For strain 5977, there was an antagonistic interaction with imidacloprid. It was difficult to obtain <30% mortality for imidacloprid only treatments, which was considered the upper limit for sublethal doses. The mean percentage mortality caused by imidacloprid was 37.5%, and this high percentage made it difficult for any combination treatment to cause significantly more mortality than the expected additive effect. These results clarify the interactions of other control products with B. bassiana and provide a basis for a reduced pesticide approach to mole cricket control.     

Euryconema brevicauda n. sp. (Oxyurida: Thelastomatidae) a parasite of the mole cricket Neocurtilla claraziana (Orthoptera: Gryllotalpidae) in Argentina

Euryconema brevicauda n. sp. parasitizing the mole cricket Neocurtilla claraziana found in Buenos Aires province, Argentina, is described and illustrated. This species is characterized by the male having 3 pairs of genital papillae, 1 pair preanal and 2 pairs postanal, and a short, conical-shaped tail.     

Biological control of the black vine weevil, Otiorhynchus sulcatus (Coleoptera: Curculionidae) with entomopathogenic nematodes (Nematoda: Rhabditida)

Trials conducted under glasshouse conditions showed that control of Otiorhynchus sulcatus larvae in strawberry plants can be effective using Steinernema carpocapsae and Heterorhabditis megidis, given that temperature and moisture extremes are avoided. In field experiments, the double line T-Tape® drip irrigation system performed better than the single line T-Tape® system, effectively distributing the nematodes along and across strawberry raised beds, and placing them close to the root zone where O. sulcatus larvae feed. As soil temperatures are satisfactory for nematode infectivity from late spring to early autumn, nematode applications were aimed at late instar larvae during spring, and early instar larvae during summer. Late summer field treatment with S. carpocapsae induced 49.5% reduction of the early instar larvae, and field application of the same nematode species in late spring resulted in 65% control of late instar larvae. In the same trial, spring application of H. megidis caused 26% mortality of late instar larvae of O. sulcatus.     

Anhydrobiotic potential and long-term storage of entomopathogenic nematodes (Rhabditida: Steinernematidae)

Anhydrobiosis is considered to be an important means of achieving storage stability of entomopathogenic nematodes that are used in biological control. This study explored the effects of anhydrobiosis on longevity and infectivity of infective juveniles (IJs) of three species of entomopathogenic nematodes Steinernema carpocapsae, Steinernema feltiae, and Steinernema riobrave at 5 and 25 degrees C. Anhydrobiosis was induced in water-dispersible granules (WG) at 0.966-0.971 water activity and 25 degrees C following a 7-day preconditioning of IJs at 5 degrees C in tap water. Survival and infectivity of the desiccated (anhydrobiotic) IJs was compared with non-desiccated IJs stored in water for different periods. Anhydrobiosis increased longevity of S. carpocapsae IJs by 3 months and of S. riobrave by 1 month in WG at 25 degrees C as compared with IJs stored in water. However, desiccation decreased S. feltiae longevity at 25 degrees C and of all three species at 5 degrees C. These results demonstrate a shelf-life of 5 months for S. carpocapsae at 25 degrees C and 9 months at 5 degrees C in WG with over 90% IJ survival. For S. feltiae, over 90% survival occurred only for 2 months at 25 degrees C and 5 months at 5 degrees C in WG. Steinernema riobrave had over 90% survival only for 1 month at 25 degrees C and the survival dropped below 85% within 1 month at 5 degrees C. Induction of anhydrobiosis in WG resulted in 85, 79 and 76% reduction in oxygen consumption by S. carpocapsae, S. feltiae, and S. riobrave IJs, respectively. Differences in IJ longevity among three species in water at 25 degrees C were related both to the initial lipid content and the rate of lipid utilisation, but not at 5 degrees C. The one-on-one infection bioassays indicated that desiccation had no negative effect on the infectivity of any of the nematode species suggesting no harmful effect on the IJs and/or their symbiotic bacteria. The species differences in IJ longevity and desiccation survival at different temperatures are discussed in relation to their foraging strategy and temperature adaptation.     

Biological control potentials of insect-parasitic nematode Rhabditis blumi (Nematoda: Rhabditida) for major cruciferous vegetable insect pests

Pathogenicity of Rhabditis blumi Sudhaus against major cruciferous insect pests was evaluated in the lab and greenhouse. In Petri-dish tests against the insects, including Artogeia rapae L., Mamestra brassicae L., and Plutella xylostella L., insect mortality by R. blumi and its associated bacteria was dose and time dependent, which increased with dose (0?C80 dauer juveniles/larva) and time increments. Pathogenicity against fourth-instar larvae was higher than the rate of corresponding third-instar larvae. The highest insect mortality rate was observed in fourth-instar larvae of P. xylostella, followed by A. rapae, and M. brassicae, with mortality rates of 93.5, 88.2, and 77.8?%, respectively. Lethal dose values at 50?% (LD₅₀) of R. blumi were 25.7 dauer juveniles/larva on P. xylostella; 28.0 dauer juveniles/larva on A. rapae; and 40.6 dauer juveniles/larva on M. brassicae, respectively. In greenhouse tests, P. xylostella larvae were most susceptible to nematodes, with insect reduction rate of 88.0?%. The rate varied with vegetable species and persistence time of live nematodes on vegetable leaves after spraying. Nematodes established in cadavers showed positive correlation with nematode dose, whereas nematode persistence on the leaf was inversely related to hours after treatment.     

Virulence of entomopathogenic nematodes (Rhabditida: Steinernematidae, Heterorhabditidae) against Tipula paludosa (Diptera: Tipulidae), a turfgrass pest on golf courses

The European crane fly (ECF), Tipula paludosa Meigen feeds on leaves, crowns, and roots of cool-season turfgrasses causing damage to residential lawns and golf courses. A laboratory study was conducted to determine the susceptibility of ECF larvae to four commercial entomopathogenic nematode (EPN) species (Heterorhabditis marelatus, H. megidis, Steinernema carpocapsae and S. feltiae). The virulence of four S. feltiae isolates recovered from golf courses in Quebec and Ontario were also compared to a commercial strain. LC₅₀ values of EPN against late instar ECF larvae were 152, 562, 763, and 3584 for S. feltiae, H. megidis, H. marelatus and S. carpocapsae, respectively. When non-feeding (without grass seedling), ECF larvae mortalities decreased for all nematode species and concentrations tested. At 25°C, LC₅₀ values for the two most virulent indigenous S. feltiae were 129 and 187 nematodes/larva, not different from the commercial strain. At 5°C, the commercial S. feltiae was more effective than both BIC14A and RE6A isolates against ECF larvae. However, at 15°C, BIC14A was the most virulent at the low concentration of 200 IJs/larva.     

Selection of insect parasitic nematodes for biological control of the garden chafer,Phyllopertha horticola

Larvae ofPhyllopertha horticola L. (Coleoptera: Scarabaeidae) cause increasing problems on sports fields and lawns in NW-Europe. A biological control programme using insect parasitic nematodes is being developed. This paper contains the results of bioassays with various species and isolates of the nematode generaHeterorhabditis andSteinernema. In bioassays in small pots with moist sand, most of the nematode isolates gave 30–60% mortality against each of the three larval stages. The susceptibility of the grubs for nematode infection generally increased with larval development.H. bacteriophora, H. heliothidis, H. megidis, a DutchHeterorhabditis isolate NLH-E87.3 andS. glaseri 326 showed the highest mortality rates, with nearly 100% mortality of third instar grubs. The DutchHeterorhabditis isolate NLH-E87.3 andS. glaseri 326 were selected as candidates for further studies on their potential as biological control agents forP. horticola grubs in the field.     

Laboratory infection of terrestrial isopods (Crustacea: Isopoda) with neoaplectanid and heterorhabditid nematodes (Rhabditida: Nematoda)

Under laboratory conditions, the nematodes Neoaplectana carpocapsae and Heterorhabditis heliothidis were able to infect and develop inside the hemocoel of terrestrial isopods belonging to the genus Porcellio. Armidillidium vulgare was also infected by N. carpocapsae, but this host was less susceptible than Porcellio. Results show that, under suitable ecological conditions, it would be feasible to utilize N. carpocapsae as a biological control agent of sowbugs (Porcellio spp.). This is the first report showing the ability of neoaplectanids and heterorhabditids to invade representatives of the Crustacea.     

Temporal association of entomopathogenic nematodes (Rhabditida: Steinernematidae and Heterorhabditidae) and bacteria

Galleria mellonella L. larvae were infected with three species (seven strains) of Steinernema spp. or three species (three strains) of Heterorhabditis spp. Infected larvae were incubated at 22, 27, and 32 degrees C. Larvae were dorsally dissected every 6h over a 48-h period. Hemolymph was collected and streaked on tryptic soy agar plates. Several non-symbiotic bacterial species were identified from infected insect cadavers: Enterobacter gergoviae, Vibrio spp., Pseudomonas fluorescens type C, Serratia marcescens, Citrobacter freundii, and Serratia proteomaculans. At 18-24 h incubation, the nematode-associated symbiont occurred almost exclusively. Bacterial associates generally appeared outside the 18-24 h window. Infective juveniles of Steinernema feltiae (Filipjev) (27), Steinernema riobrave Cabanillas, Poinar, and Raulston (Oscar), or Steinernema carpocapsae (Weiser) (Kapow) were left untreated, or surface sterilized using thimerosal, then pipetted under sterile conditions onto tryptic soy agar plates. Several additional species of associated bacteria were identified using this method compared with the less extensive range of species isolated from infected G. mellonella. There was no difference in bacterial species identified from non-sterile or surface sterilized nematodes, suggesting that the bacteria identified originated from either inside the nematode or between second and third stage juvenile cuticles. Infective juveniles of S. feltiae (Cowles), S. carpocapsae (Cowles), and H. bacteriophora Poinar (Cowles) were isolated from field samples. Nematodes were surface-sterilized using sodium hypochlorite, mixed with G. mellonella hemolymph, and pipetted onto Biolog BUG (with blood) agar. Only the relevant symbionts were isolated from the limited number of samples available. The nematodes were then cultured in the laboratory for 14 months (sub-cultured in G. mellonella 7-times). Other Enterobacteriaceae could then be isolated from the steinernematid nematodes including S. marcescens, Salmonella sp., and E. gergoviae, indicating the ability of the nematodes to associate with other bacteria in laboratory culture.     

Biological control: a novel strategy for the control of the plant parasitic nematodes

Antonie van Leeuwenhoek - Plant parasitic nematodes (Root-knot nematodes, Meloidogyne spp.) are rounded worms, microscopic, and cause many agricultural economic losses. Their attacks have a direct...     

Diversity and distribution of entomopathogenic nematodes (Rhabditida: Steinernematidae and Heterorhabditidae) in Turkey

The diversity and distribution of entomopathogenic nematodes in thefamilies Steinernematidae and Heterorhabditidae were assessed throughout anextensive soil survey in Turkey during 1999 and 2000. Entomopathogenic nematodeswere recovered from six out of seven regions sampled, with 22 positive sites(2%) out of 1080 sites sampled. A single nematode isolate was recovered at eachof the positive sites, of which 15 were steinernematid isolates and seven wereheterorhabditid isolates representing a total of four species. Based onmorphometric and molecular data, the nematode species were identified asHeterorhabditis bacteriophora, Steinernemafeltiae, S. affine, andSteinernema n. sp. The most common species was S.feltiae, which was isolated from 10 sites in six regions, followed byH. bacteriophora from seven sites in five regions,S. affine from four sites in two regions, andSteinernema n. sp. from one site. Heterorhabditisbacteriophora and S. feltiae have been found inmany parts of the world, whereas S. affine, so far, hasonly been recovered in Europe until our survey. Steinernemaaffine was isolated from the European (Marmara) as well as theAsiatic region (Middle Anatolia) of Turkey. A new undescribedSteinernema sp. was isolated from the most eastern region(East Anatolia) of Turkey. Soils of the positive sites were classified as sandy,sandy loam, or loam (68.2%) and sandy–clay–loam or clay loam (31.8%) and the pHranged from 5.6 to 7.9. The habitats from which the entomopathogenic nematodeswere isolated were broadly classified as disturbed (59.1%), which includedagricultural fields and poplar planted for lumber and wind breaks, andundisturbed (40.9%), which included pine forest, grassland, marsh and reed sites.Steinernema feltiae, S. affine, andH. bacteriophora were recovered from both disturbed andundisturbed habitats. The new Steinernema sp. was recoveredfrom grassland. Our survey showed that these nematodes occur widely throughoutTurkey, but at a frequency below that reported for other parts of the world.     

New species of mole crickets of the Gryllotalpa gryllotalpa group (Orthoptera: Gryllotalpidae) from Israel, based on morphology, song recordings, chromosomes and cuticular hydrocarbons, with comments on the distribution of the group in Europe and the Mediterranean region

Two sibling species of mole crickets of the Gryllotalpa gryllotalpa group inhabit Israel. Both are described as new species based on differences in their morphology, acoustic behaviour, chromosome number, cuticular hydrocarbon pattern and habitats. The species with males having 2n = 23 chromosomes, described in the past as the 'Dead Sea race', is designated as G. marismortui sp.n. It is an endangered species. In morphology and chromosome number it is similar to G. cossyrensis Bacetti & Capra from Italy, but differs in the composition of cuticular hydrocarbons. Gryllotalpa marismortui occurs in a limited geographical area along the shore of the Dead Sea. Another species, with 2n = 19 chromosomes, is described as G. tali sp.n. and is found throughout Israel. The calling songs of the males of the new species differ markedly. Females of the respective chromosomal numbers discriminate in favour of the homospecific call. Their habitats are strikingly different: G. marismortui lives in hypersaline soil along the Dead Sea shore (total chloride 187.25 meq %), G. tali inhabits freshwater soil (9.31 meq %). The distribution of the twelve known sibling species and one chromosomal race (not yet described) of the G. gryllotalpa group from Europe and the eastern Mediterranean is summarized. The zoogeography and phylogeny of the group are discussed.